COMPOSITIONS AND METHODS OF NUCLEIC ACID PREPARATION AND ANALYSES

摘要

The present invention provides methods of generating single-stranded polynucleotides comprising use of adaptor sequence(s), single-stranded polynucleotide amplification and a primer comprising BJSA. Methods of analysing one or more regions on a desired polynucleotide using probes and single-stranded polynucleotides are also provided. Also provided are kits and compositions useful for these methods.

主权项

1. A method of generating single-stranded polynucleotides comprising asymmetric adaptor sequences, comprising:
i) ligating one or more DNA fragments to: a) a first adaptor comprising a tag; and b) a second adaptor comprising a recognition sequence; ii) selecting DNA fragments comprising the first adaptor based on the presence of the tag; iii) amplifying the DNA fragments selected from step ii) by single-strand polynucleotide amplification using a primer comprising RNA and hybridizing the primer comprising RNA to the recognition sequence, thereby selectively amplifying DNA fragments comprising the second adaptor to obtain a population of single-stranded polynucleotides.